Injectable ready-to-use solutions containing an antitumor anthracycline glycoside

ABSTRACT

According to the invention there is provided a sterile, pyrogen-free, ready-to-use solution of an anthracycline glycoside, especially doxorubicin, which consists essentially of a physiologically acceptable salt of an anthracycline glycoside dissolved in a physiologically acceptable solvent therefor, which has not been reconstituted from a lyophilizate and which has a pH of from 2.5 to 6.5. The solution of the invention is particularly advantageous for the administration by injection of the anthracycline glycoside drugs, e.g. doxorubicin, in the treatment of both human and animal tumors.

This is a division of application Ser. No. 07/503,856, filed on Apr. 3,1990, now U.S. Pat. No. 5,124,317.

The present invention relates to a stable intravenously injectableready-to-use solution of an antitumor anthracycline glycoside, e.g.doxorubicin, to a process for preparing such a solution, and provide thesame in a sealed container, and to a method for treating tumors by theuse of the said ready-to-use solution.

The anthracycline glycoside compounds are a well known class ofcompounds in the antineoplastic group of agents, wherein doxorubicin isa typical, and the most widely used, representative: Doxorubicin.Anticancer Antibiotics, Federico Arcamone, 1981, Publ: Academic Press,New York, N.Y.; Adriamycin Review, EROTC International Symposium,Brussels, May, 1974, edited by M.Staquet, Publ. Eur. Press Medikon,Ghent, Belg.;

Results of Adriamycin Therapy, Adriamycin Symposium at Frankfurt/Main1974 edited by M. Ghione, J. Fetzer and H. Maier, publ.: Springer, NewYork, N.Y.

At present, anthracycline glycoside antitumor drugs, in particular,e.g., doxorubicin, are solely available in the form of lyophilizedpreparations, which need to be Reconstituted before administration.

Both the manufacturing and the reconstitution of such preparationsexpose the involved personnel (workers, pharmacists, medical personnel,nurses) to risks of contamination which are particularly serious due tothe toxicity of the antitumor substances.

The Martindale Extra Pharmacopoeia 28th edition, page 175 left column,reports, indeed, about adverse effects of antineoplastic drugs andrecommends that "They must be handled with great care and contact withskin and eyes avoided; they should not be inhaled. Care must be taken toavoid extravasation since pain and tissue damage may ensue."

Similarly, Scand. J. Work Environ Health vol.10 (2), pages 71-74 (1984),as well as articles on Chemistry Industry, Issue Jul. 4, 1983, page 488,and Drug-Topics-Medical-Economics-Co, Issue Feb. 7, 1983, page 99 reportabout severe adverse effects observed in medical personnel exposed touse of cytostatic agents, including doxorubicin.

To administer a lyophilized preparation, double handling of the drug isrequired, the lyophilized cake having to be first reconstituted and thenadministered and,moreover,in some cases, the complete dissolution of thepowder may require prolonged shaking because of solubilization problems.

As the risks connected with the manufacturing and the reconstitution ofa lyophilized preparate would be highly reduced if a ready-to-usesolution of the drug were available, we have developed a stable,therapeutically acceptable intravenously injectable solution of ananthracycline glycoside drug, e.g. doxorubicin, whose preparation andadministration does not require either lyophilization or reconstitution.

According to the present invention, there is provided a sterile,pyrogen-free, anthracycline glycoside solution which consistsessentially of a physiologically acceptable salt of an anthracyclineglycoside dissolved in a physiologically acceptable solvent therefor,which has not been reconstituted from a lyophilizate and which has a pHof from 2.5 to 6.5.

Preferably the solution of the invention is provided in a sealedcontainer.

Preferably the anthracycline glycoside is chosen from the groupconsisting of doxorubicin, 4'-epi-doxorubicin (i.e. epirubicin),4'-desoxy-doxorubicin (i.e. esorubicin), 4'-desoxy-4'-iodo-doxorubicin,daunorubicin and 4-demethoxydaunorubicin (i.e. idarubicin).

A particularly preferred anthracycline glycoside is doxorubicin.

Any physiologically acceptable salt of the anthracycline glycoside maybe used for preparing the solution of the invention. Examples ofsuitable salts may be, for instance, the salts with mineral inorganicacids such as hydrochloric, hydrobromic, sulfuric, phosphoric, nitricand the like, and the salts with certain organic acids such as acetic,succinic, tartaric, ascorbic, citric, glutammic, benzoic,methanesulfonic, ethanesulfonic and the like. The salt with hydrochloricacid is a particularly preferred salt, especially when the anthracyclineglycoside is doxorubicin.

Any solvent which is physiologically acceptable and which is able todissolve the anthracycline glycoside salt may be used. The solution ofthe invention may also contain one or more additional components such asa co-solubilizing agent (which may be the same as a solvent), a tonicityadjustment agent and a preservative. Examples of solvents,co-solubilizing agents, tonicity adjustment agents and preservativeswhich can be used for the preparation of the anthracycline glycosidesolutions of the invention are hereunder reported.

Suitable solvents and co-solubilizing agents may be, for instance,water; physiological saline; aliphatic amides, e.g.N,N-dimethylacetamide, N-hydroxy-2-ethyl-lactamide and the like;alcohols, e.g. ethanol, benzyl alcohol and the like; glycols andpolyalcohols, e.g. propyleneglycol, glycerin and the like; esters ofpolyalcohols, e.g. diacetine, triacetine and the like; polyglycols andpolyethers, e.g. polyethyleneglycol 400, propyleneglycol methylethersand the like; dioxolanes, e.g. isopropylidenglycerin and the like;dimethylisosorbide; pyrrolidone derivatives, e.g. 2-pyrrolidone,N-methyl-2-pyrrolidone, polyvinylpyrrolidone (co-solubilizing agentonly) and the like; polyoxyethylenated fatty alcohols, e.g. Brij^(R) andthe like; esters of polyoxyethylenated fatty acids, e.g. Cremophor^(R),Myrj^(R) and the like; polysorbates, e.g. Tweens^(R) ; polyoxyethylenederivatives of polypropyleneglycols, e.g. Pluronics^(R).

A particularly preferred co-solubilizing agent is polyvinylpyrrolidone.

Suitable tonicity adjustment agents may be, for instance,physiologically acceptable inorganic chlorides, e.g. sodium chloride,dextrose, lactose, mannitol and the like.

Preservatives suitable for physiological administration may be, forinstance, esters of para-hydroxybenzoic acid (e.g., methyl, ethyl,propyl and butyl esters, or mixtures of them), chlorocresol and thelike.

The above mentioned solvents and co-solubilizing agents, tonicityadjustment agents and preservatives can be used alone or as a mixture oftwo or more of them.

Examples of preferred solvents are water, ethanol, polyethyleneglycoland dimethylacetamide as well as mixtures in various proportions ofthese solvents. Water is a particularly preferred solvent.

To adjust the pH within the range of from 2.5 to about 5.0 aphysiologically acceptable acid may be added as desired. The acid may beany physiologically acceptable acid, e.g., an inorganic mineral acidsuch as hydrochloric, hydrobromic, sulfuric, phosphoric, nitric and thelike, or an organic acid such as acetic, succinic, tartaric, ascorbic,citric, glutammic, benzoic, methanesulphonic, ethanesulfonic and thelike, or also an acidic physiologically acceptable buffer solution,e.g., a chloride buffer, an acetate buffer, a phosphate buffer and thelike.

For obtaining pH values from about 5 to about 5.5 the addition of theacid is not, usually, necessary, but only addition of a physiologicallyacceptable buffer solution, e.g., one of those indicated above, may berequired, as desired.

For obtaining pH values from about 5.5 to 6.5 the addition of aphysiologically acceptable alkalinizing agent, such as sodium hydroxide,a mono, di- or triethanolamine or the like, or, preferably, buffersolution such as a phosphate buffer, a TRIS buffer or the like isrequired.

The preferred range of pH for the ready-to-use solution of the inventionis from 2.5 to 5.5, in particular from about 3 to about 5.2, a pH ofabout 3 and a pH of about 5 being particularly preferred values.

In the solutions of the invention the concentration of the anthracyclineglycoside may vary within broad ranges, preferably, from 0.1 mg/ml to100 mg/ml, in particular from 0.1 mg/ml to 50 mg/ml, most preferablyfrom 1 mg/ml to 20 mg/ml.

The preferred ranges of concentration may be slightly different fordifferent anthracycline glycosides. Thus, for example, preferredconcentrations for doxorubicin are from about 2 mg/ml to about 50 mg/ml,preferably from 2 mg/ml to 20 mg/ml, particularly appropriate valuesbeing 2 mg/ml and 5 mg/ml. Similar concentrations are preferred also for4'-epi-doxorubicin, 4'-desoxy-doxorubicin and4'-desoxy-4'-iodo-doxorubicin. Preferred ranges of concentration fordaunorubicin and 4-demethoxy-daunorubicin are from 0.1 mg/ml to 50mg/ml, preferably from 1 mg/ml to 20 mg/ml, concentrations of 1 mg/mland 5 mg/ml being particularly appropriate.

Suitable packaging for the anthracycline glycoside solutions may be allapproved containers intended for parenteral use, such as plastic andglass containers, ready-to-use syringes and the like. Preferably thecontainer is a sealed glass container, e.g. a vial or an ampoule.

According to a particularly preferred feature of the invention, there isprovided a sterile, pyrogen-free, doxorubicin solution which consistsessentially of a physiologically acceptable salt of doxorubicindissolved in a physiologically acceptable solvent therefor, which hasnot been reconstituted from a lyophilizate and which has a pH of from2.5 to 6.5.

In the above indicated preferred feature of the invention thephysiologically acceptable salt of doxorubicin may be, e.g. the saltwith a mineral inorganic acid such as hydrochloric, hydrobromic,sulfuric, phosphoric, nitric and the like, or the salt with an organicacid such as acetic, succinic, tartaric, ascorbic, citric, glutammic,benzoic, methanesulfonic, ethanesulfonic and the like. The hydrochloridesalt is a particularly preferred salt.

For the solution hereabove indicated as a preferred feature of theinvention suitable solvents, co-solubilizing agents, tonicity adjustmentagents and preservatives may be the same as those previously recited inthis specification. Water is a particularly preferred solvent.

Also, the physiologically acceptable acid which may be added to adjustthe pH to from 2.5 to about 5, if desired, and the alkanilizing agentwhich may be added to adjust the pH, if desired, to a value from about5.5 to 6.5 may be one of those previously specified. When it is desiredto adjust the pH of the above said preferred solution to a value of from2.5 to about 5, hydrochloric acid is an especially preferred acid.Preferred pH values for the above said preferred solutions of theinvention are from 2.5 to 5.5, in particular from about 3 to about 5.2,the pH values of 3 and 5 being especially preferred.

Though the concentration of doxorubicin in the above preferred featuremay vary within the broad range from 0.1 mg/ml to 100 mg/ml, preferredconcentrations are from 2 mg/ml to 50 mg/ml, most preferably from 2mg/ml to 20 mg/ml: examples of especially preferred concentrations ofdoxorubicin are 2 mg/ml and 5 mg/ml.

The invention also provides a process for producing a sterile,pyrogen-free anthracycline glycoside solution with a pH of from 2.5 to6.5, which process comprises dissolving a physiologically acceptablesalt of the anthracycline glycoside, which salt is not in the form of alyophilizate, in a physiologically acceptable solvent therefor;optionally adding a physiologically acceptable acid or buffer to adjustthe pH within the said range as desired; and passing the resultingsolution through a sterilising filter.

One or more additional components such as co-solubilizing agents,tonicity adjustment agents and preservatives, for instance of the kindpreviously specified, may be added to the solution prior to passing thesolution through the sterilising filter.

With the solutions of the invention it is possible to obtaincompositions having a very high concentration of the anthracyclineglycoside active substance even at 50 mg/ml and more. This constitutes agreat advantage over the presently available lyophilized preparateswherein high concentrations of anthracycline glycoside can only beobtained with difficulty because of solubilization problems encounteredin reconstitution, mainly with saline. The presence of the excipient,e.g. lactose, in the lyophilized cake, and its generally high proportionin respect of the active substance, even up to 5 parts of excipient perpart of active substance, has a negative effect on solubilization sothat difficulties may arise in obtaining dissolution of the lyophilizedcake, especially for concentrations of anthracycline glycoside higherthan 2 mg/ml.

The solutions of the invention are characterized by a good stability.Solutions in various solvents and with different pH's and concentrationshave been found to be stable for long periods at temperatures acceptedfor the storage of pharmaceutical preparations. This is illustrated inthe Examples which follow.

Owing to the well known anti-tumor activity of the anthracyclineglycoside active drug substance, the pharmaceutical compositions of theinvention are useful for treating tumors in both human and animal hosts.Examples of tumors that can be treated are, for instance, sarcomas,including osteogenic and soft tissue sarcomas, carcinomas, e.g.,breast-, lung-, bladder-, thyroid-, prostate- and ovarian carcinoma,lymphomas, including Hodgkin and non-Hodgkin lymphomas, neuroblastoma,melanoma, myeloma, Wilms tumor, and leukemias, including acutelymphoblastic leukemia and acute myeloblastic leukemia. Examples ofspecific tumours that can be treated are Moloney sarcoma Virus, Sarcoma180 Ascites, solid Sarcoma 180, gross transplantable leukemia, L 1210leukemia and lymphocytic P 388 leukemia. Thus, according to theinvention there is also provided a method of inhibiting the growth of atumour, in particular one of those indicated above, which comprisesadministering to a host suffering from said tumour an injectablesolution according to the invention containing the active drug substancein an amount sufficient to inhibit the growth of said tumour. Theinjectable solutions of the invention are administered b rapidintravenous injection or infusion according to a variety of possibledose schedules. Suitable dose schedule for doxorubicin may be, forexample, of 60 to 75 mg of active drug substance per m² of body surfacegiven as a single rapid infusion and repeated at 21 days; an alternativeschedule may be of 30 mg/m² day by intravenous route for 3 days, every25 days. Suitable dosages for 4'-epi-doxorubicin and 4'-desoxy-doxorubicin may be, for instance, of 75 to 90 mg/m² given in asingle infusion to be repeated at 21 days, and similar dosages may beuseful also for 4 '-desoxy-4 '-iodo-doxorubicin. Idarubicin, i.e.4-demethoxy-daunorubicin, may be. e.g., administered intravenously at asingle dose of 13-15 mg//m² ever, 21 days in the treatment of solidtumours, while in the treatment of leukemias a preferred dose scheduleis, e.g., of 10-12 mg/m² day by intravenous route for 3 days, to berepeated every 15-21 days; similar dosages may be, e.g., followed alsofor daunorubicin.

The following examples illustrate but do not limit in any way theinvention.

With reference to the examples,the stability controls on theready-to-use solutions were carried out by means of high performanceliquid chromatography (HPLC), at the following

    ______________________________________                                        Liquid chromatograph                                                                            Varian model 5010                                           Spectrophotometric detector                                                                     Knauer model 8700                                           Integrating recorder                                                                            Varian model CDS 401                                        Injection valve   Rheodyne model 7125 fitted with                                               a 10 mcl sample loop                                        Chromatographic column                                                                          Waters μ-Bondapak C18                                                      (length = 300 mm; inner diameter                                              = 3.9 mm; average particle size =                                             10 mcm)                                                     Column temperature                                                                              ambient (about 22° C. ± 2° C.)             Mobile phase      water : acetonitrile (69:31 v/v)                                              adjusted to pH 2 with phosphoric                                              acid, filtered (sintered glass                                                filter, 1 mcm or finer porosity)                                              and deaerated                                               Mobile phase flow rate                                                                          1.5 ml/min                                                  Analytical wavelength                                                                           254 ± 1 nm                                               Integrating recorder sensitivity                                                                512                                                         Chart speed       1 cm/min                                                    ______________________________________                                    

At these conditions, the peak of the anthracycline glycoside showed aretention time of about 6 minutes. The obtained results are reported inthe Tables accompanying the examples.

The extrapolation of the analytical data in order to determine the timewhen the 90% of the initial assay could be expected (t₉₀ value) was madefollowing an Arrhenius plot.

This procedure of analytical data treatment is well known and widelyused and described in the art: see, e.g., Chemical Stability ofPharmaceuticals, Kennet A. Connors, Gordon L. Amidon, Lloyd Kennon,Publ. John Wiley and Sons, New York, N.Y., 1979.

The term "teflon" refers to "Teflon™".

EXAMPLE 1

    ______________________________________                                        Composition     for 80 vials (for 1 vial)                                     ______________________________________                                        Doxorubicin.HCl 0.8 g        (10 mg)                                          Water for injections                                                                          0.4 l        (5 ml)                                           q.s. to                                                                       ______________________________________                                    

Doxorubicin.HCl (0.80 g) was dissolved in 90 percent of the amount ofwater for injections, de-aerated by nitrogen bubbling. The pH of thesolution was not adjusted. Further de-aerated water for injections wasthen added to bring the solution to its final volume (0.40 1).

The solution was filtered through a 0.22μ microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (550° C.), 4 weeks(45° C. and 35° C.) and 12 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 1:

                                      TABLE 1                                     __________________________________________________________________________    INITIAL VALUES                                                                Concentration:1.994 mg/ml                                                                      pH = 5.2                                                     Relative % Assay: 100.0                                                       TEMPERATURE                                                                   4° C. 35° C.                                                                          45° C.                                                                         55° C.                                   TIME Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                                                              Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                       (weeks)                                                                            mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                                                              mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                       __________________________________________________________________________    1    1.992                                                                             99.9                                                                              1.917                                                                             96.1 1.768                                                                             88.7                                                                              1.493                                                                             75.0                                        2            1.843                                                                             92.4 1.618                                                                             81.1                                                                              1.166                                                                             58.5                                        3            1.774                                                                             89.0 1.506                                                                             75.5                                                                              0.830                                                                             41.6                                        4    1.974                                                                             99.0                                                                              1.720                                                                             86.3 1.393                                                                             69.9                                                12   1.980                                                                             99.3                                                                 __________________________________________________________________________     t.sub.90 (days) extrapolated according to Arrhenius equation:                 t.sub.90 at 4° C. = 815 days                                           t.sub.90 at 8° C. = 480 days                                      

Similar stability data can be observed also for analogous solutionscontaining either doxorubicin hydrochloride at 5 mg/ml concentration, or4'-epi-doxorubicin, 4'-desoxy-doxorubicin,4'-desoxy-4'-iodo-doxorubicin, daunorubicin or 4-demethoxy-daunorubicin,as hydrochloride salts, at both 2 mg/ml and 5 mg/ml concentration.

EXAMPLE 2

    ______________________________________                                        Composition       for 80 vials                                                                              (for 1 vial)                                    ______________________________________                                        Doxorubicin.HCl   0.8 g       (10 mg)                                         Hydrochloric acid 0.1N                                                                          pH = 3      (pH = 3)                                        q.s. to                                                                       Water for injections                                                                            0.4 l       (5 ml)                                          q.s. to                                                                       ______________________________________                                    

Doxorubicin.HCl (0.8 g) was dissolved in 90 percent of the amount ofwater for injections, de-aerated by nitrogen bubbling. The hydrochloricacid was then added dropwise to adjust the pH of the solution to. 3.Further de-aerated water for injections was then added to bring thesolution to its final volume (0.4 1).

The solution was filtered through a 0.22μ microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 12 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 2:

                                      TABLE 2                                     __________________________________________________________________________    INITIAL VALUES                                                                Concentration:1.992 mg/ml                                                                      pH = 3.0                                                     Relative % Assay: 100.0                                                       TEMPERATURE                                                                   4° C. 35° C.                                                                          45° C.                                                                         55° C.                                   TIME Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                                                              Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                       (weeks)                                                                            mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                                                              mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                       __________________________________________________________________________    1    1.995                                                                             100.2                                                                             1.952                                                                             98.0 1.919                                                                             96.3                                                                              1.493                                                                             75.0                                        2            1.889                                                                             94.8 1.851                                                                             92.9                                                                              1.036                                                                             51.9                                        3            1.876                                                                             94.2 1.565                                                                             78.6                                                                              0.730                                                                             36.7                                        4    1.979                                                                             99.4                                                                              1.808                                                                             90.8 1.393                                                                             69.9                                                12   1.972                                                                             99.0                                                                 __________________________________________________________________________     t.sub.90 (days) extrapolated according to Arrhenius equation:                 t.sub.90 at 4° C. = 3970 days                                          t.sub.90 at 8° C. = 2000 days                                     

Similar stability data can be observed also for analogous solutionscontaining either doxorubicin hydrochloride at 5 mg/ml concentration, or4'-epi-doxorubicin, 4'-desoxy-doxorubicin,4'-desoxy-4'-iodo-doxorubicin, daunorubicin or 4-demethoxy-daunorubicin,as hydrochloride salts, at both 2 mg/ml and 5 mg/ml concentration.

EXAMPLE 3

    ______________________________________                                        Composition       for 80 vials                                                                              (for 1 vial)                                    ______________________________________                                        Doxorubicin.HCl   8.0 g       (100 mg)                                        Hydrochloric acid 0.1N                                                                          pH = 3      (pH = 3)                                        q.s. to                                                                       Water for injections                                                                            0.4 l       (5 ml)                                          q.s. to                                                                       ______________________________________                                    

Doxorubicin.HCl (8.0 g) was dissolved in 90 percent of the amount ofwater for injections, de-aerated by nitrogen bubbling. The hydrochloricacid was then added dropwise to adjust the pH of the solution to 3.Further de-aerated water for injections was then added to bring thesolution to its final volume (0.4 1).

The solution was filtered through a 0.22μ microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 12 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 3:

                                      TABLE 3                                     __________________________________________________________________________    INITIAL VALUES                                                                Concentration:20.06 mg/ml                                                                      pH = 2.95                                                    Relative % Assay: 100.0                                                       TEMPERATURE                                                                   4° C. 35° C.                                                                          45° C.                                                                         55° C.                                   TIME Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                                                              Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                       (weeks)                                                                            mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                                                              mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                       __________________________________________________________________________    1    20.06                                                                             100.0                                                                             19.56                                                                             97.5 17.84                                                                             88.9                                                                              12.31                                                                             61.4                                        2            18.87                                                                             94.1 15.61                                                                             77.8                                                                              7.09                                                                              35.3                                        3            18.24                                                                             90.9 13.41                                                                             66.8                                                                              3.13                                                                              15.6                                        4    19.91                                                                             99.2                                                                              17.51                                                                             87.3 11.07                                                                             55.2                                                12   19.80                                                                             98.7                                                                 __________________________________________________________________________     t.sub.90 (days) extrapolated according to Arrhenius equation:                 t.sub.90 at 4° C. = 3700 days                                          t.sub.90 at 8° C. = 1780 days                                     

Similar stability data can be observed for analogous solutionscontaining 4'-epi-doxorubicin or 4'-desoxy-doxorubicin, as hydrochloridesalts, at the same 20 mg/ml concentration.

EXAMPLE 4

    ______________________________________                                        Composition      for 80 vials                                                                              (for 1 vial)                                     ______________________________________                                        Doxorubicin.HCl  0.80 g      (10.0 mg)                                        Polyvinylpyrrolidone                                                                           20.00 g     (250.0 mg)                                       Water for injections                                                                           0.40 l      (5.0 ml)                                         q.s. to                                                                       ______________________________________                                    

Doxorubicin.HCl (0.80 g) was dissolved in 90 percent of the amount ofwater for injections, de-aerated by nitrogen bubbling. The pH of thesolution was not adjusted. Polyvinylpyrrolidone was added and dissolvedunder stirring and nitrogen bubbling. Further de-aerated water forinjections was then added to bring the solution to its final volume(0.40 1).

The solution was filtered through a 0.22μ microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 8 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 4:

                                      TABLE 4                                     __________________________________________________________________________    INITIAL VALUES                                                                Concentration:1.986 mg/ml                                                                      pH = 4.6                                                     Relative % Assay: 100.0                                                       TEMPERATURE                                                                   4° C. 35° C.                                                                          45° C.                                                                         55° C.                                   TIME Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                                                              Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                       (weeks)                                                                            mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                                                              mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                       __________________________________________________________________________    1    1.984                                                                             99.9                                                                              1.928                                                                             97.1 1.797                                                                             90.5                                                                              1.605                                                                             80.8                                        2            1.847                                                                             93.0 1.616                                                                             81.4                                                                              1.293                                                                             65.1                                        3            1.828                                                                             92.0 1.527                                                                             76.9                                                                              1.018                                                                             51.3                                        4    1.928                                                                             97.1                                                                              1.797                                                                             90.5 1.403                                                                             70.7                                                8    1.989                                                                             100.1                                                                __________________________________________________________________________     t.sub.90 (days) extrapolated according to Arrhenius equation:                 t.sub.90 at 4° C. = 1460 days                                          t.sub.90 at 8° C. = 835 days                                      

Similarr stability data can be observed also for analogous solutionscontaining either doxorubicin hydrochloride at 5 mg/ml concentration, or4'-epi-doxorubicin, 4'-desoxy-doxorubicin, 4'-desoxy-4'-iodo-doxorubicin, daunorubicin or 4-demethoxy-daunorubicin, ashydrochloride salts, at both 2 mg/ml and 5 mg/ml concentration.

EXAMPLE 5

    ______________________________________                                        Composition      for 80 vials                                                                              (for 1 vial)                                     ______________________________________                                        Doxorubicin.HCl  0.800 g     (10.00 mg)                                       N,N-Dimethylacetamide                                                                          0.060 l     (0.75 ml)                                        Propylene glycol 0.048 l     (0.60 ml)                                        Ethanol          0.012 l     (0.15 ml)                                        Hydrochloric acid 0.1N                                                                         pH = 3      (pH = 3)                                         q.s. to                                                                       Water for injections                                                                           0.400 l     (5.00 ml)                                        q.s. to                                                                       ______________________________________                                    

Doxorubicin.HCl (0.800 g) was dissolved in 90 percent of the amount ofwater for injections, de-aerated by nitrogen bubbling.N,N-dimethylacetamide, propylene glycol and ethanol were subsequentlyadded under stirring and nitrogen bubbling. The hydrochloric acid wasthen added dropwise to adjust the pH of the solution to 3. Furtherde-aerated water for injections was then added to bring the solution toits final volume (0.400 1).

The solution was filtered through a 0.22μ microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 8 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 5:

                                      TABLE 5                                     __________________________________________________________________________    INITIAL VALUES                                                                Concentration:2.000 mg/ml                                                                      pH = 3.03                                                    Relative % Assay: 100.0                                                       TEMPERATURE                                                                   4° C. 35° C.                                                                          45° C.                                                                         55° C.                                   TIME Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                                                              Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                       (weeks)                                                                            mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                                                              mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                       __________________________________________________________________________    1            1.892                                                                             94.6 1.735                                                                             86.7                                                                              1.495                                                                             74.7                                        2    1.993                                                                             99.7                                                                              1.927                                                                             96.4 1.624                                                                             81.2                                                                              1.212                                                                             60.6                                        3            1.908                                                                             95.4 1.432                                                                             71.6                                                                              1.032                                                                             51.6                                        4    2.00                                                                              100.0                                                                             1.863                                                                             93.2 1.266                                                                             63.3                                                8    1.960                                                                             98.0                                                                 __________________________________________________________________________     t.sub.90 (days) extrapolated according to Arrhenius equation:                 t.sub.90 at 4° C. = 4360 days                                          t.sub.90 at 8° C. = 2200 days                                     

Similar stability data can be observed also for analogous solutionscontaining either doxorubicin hydrochloride at 5 mg/ml concentration, or4'-epi-doxorubicin, 4'-desoxy-doxorubicin, 4'-desoxy-iodo-doxorubicin,daunorubicin or 4-demethoxy-daunorubicin, hydrochloride salts, at both 2mg/ml and 5 mg/ml concentration.

EXAMPLE 6

    ______________________________________                                        Composition      for 80 vials                                                                              (for 1 vial)                                     ______________________________________                                        Doxorubicin.HCl  0.8 g       (10.0 mg)                                        Polyvinylpyrrolidone                                                                           20.0 g      (250.0 mg)                                       Hydrochloric acid 0.1N                                                                         pH = 3      (pH = 3)                                         q.s. to                                                                       Water for injections                                                                           0.4 l       (5.0 ml)                                         q.s. to                                                                       ______________________________________                                    

Doxorubicin.HCl (0.8 g) was dissolved in 90 percent of the amount ofwater for injections, de-aerated by nitrogen bubbling.Polyvinylpyrrolidone was added and dissolved under stirring and nitrogenbubbling. The hydrochloric acid was then added dropwise to adjust the pHof the solution to 3. Further de-aerated water for injections was thenadded to bring the solution to its final volume (0.4 1).

The solution was filtered through a 0.22μ microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 8 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 6;

                                      TABLE 6                                     __________________________________________________________________________    INITIAL VALUES                                                                Concentration: 1.973mg/ml                                                                      pH = 2.71                                                    Relative % Assay: 100.0                                                       TEMPERATURE                                                                   4° C. 35° C.                                                                          45° C.                                                                         55° C.                                   TIME Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                                                              Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                       (weeks)                                                                            mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                                                              mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                       __________________________________________________________________________    1    2.028                                                                             102.8                                                                             1.944                                                                             98.5 1.791                                                                             90.8                                                                              1.477                                                                             74.9                                        2            1.885                                                                             95.5 1.582                                                                             80.2                                                                              0.972                                                                             49.3                                        3            1.840                                                                             93.2 1.402                                                                             71.0                                                                              0.632                                                                             32.0                                        4    1.913                                                                             97.0                                                                              1.853                                                                             93.9 1.273                                                                             64.5                                                8    1.972                                                                             99.9                                                                 __________________________________________________________________________     t.sub.90 (days) extrapolated according to Arrhenius equation:                 t.sub.90 at 4° C. = 5560 days                                          t.sub.90 at 8° C. = 2670 days                                     

Similar stability data can be observed also for analogous solutionscontaining either doxorubicin hydrochloride at 5 mg/ml concentration, or4'-epi-doxorubicin, 4'-desoxy-doxorubicin,4'-desoxy-4'-iodo-doxorubicin, daunorubicin or 4-demethoxy-daunorubicin,as hydrochloride salts, at both 2 mg/ml and 5 mg/ml concentration.

EXAMPLE 7

    ______________________________________                                        Composition      for 80 vials                                                                              (for 1 vial)                                     ______________________________________                                        Doxorubicin.HCl  8.00 g      (100.0 mg)                                       N,N-Dimethylacetamide                                                                          0.12 l      (1.5 ml)                                         Hydrochloric acid 0.1N                                                                         pH = 3      (pH = 3)                                         q.s. to                                                                       Water for injections                                                                           0.40 l      (5.0 ml)                                         q.s. to                                                                       ______________________________________                                    

Doxorubicin.HCI (8.00 g) was dissolved in 90 percent of the amount ofwater for injections, de-aerated by nitrogen bubbling.N,N-dimethylacetamide was added under stirring and nitrogen bubbling.The hydrochloric acid was then added dropwise to adjust the pH of thesolution to 3. Further de-aerated water for injections was then added tobring the solution to its final volume (0.40 1).

The solution was filtered through a 0.22μ microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 8 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 7:

                                      TABLE 7                                     __________________________________________________________________________    INITIAL VALUES                                                                Concentration:19.32 mg/ml                                                                      pH = 2.96                                                    Relative % Assay: 100.0                                                       TEMPERATURE                                                                   4° C. 35° C.                                                                          45° C.                                                                         55° C.                                   TIME Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                                                              Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                       (weeks)                                                                            mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                                                              mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                       __________________________________________________________________________    1    20.1                                                                              103.5                                                                             19.14                                                                             99.1 17.34                                                                             89.8                                                                              15.57                                                                             80.6                                        2            19.20                                                                             99.4 15.77                                                                             81.6                                                                              12.94                                                                             67.0                                        3            18.06                                                                             93.5 14.85                                                                             76.9                                                                              11.61                                                                             60.1                                        4    20.03                                                                             103.7                                                                             17.81                                                                             92.2 13.78                                                                             71.3                                                8    19.99                                                                             103.5                                                                __________________________________________________________________________     t.sub.90 (days) extrapolated according to Arrhenius equation:                 t.sub.90 at 4° C. = 1310 days                                          t.sub.90 at 8° C. = 770 days                                      

Similar stability data can be observed for analogous solutionscontaining 4'-epi-doxorubicin or 4'-desoxy-doxorubicin, as hydrochloridesalts, at the same 20 mg/ml concentration.

EXAMPLE 8

    ______________________________________                                        Composition      for 80 vials                                                                              (for 1 vial)                                     ______________________________________                                        Doxorubicin.HCl  0.80 g      (10.0 mg)                                        Ethanol          0.12 l      (1.5 ml)                                         Hydrochloric acid 0.1 N                                                                        pH = 3      (pH = 3)                                         q.s. to                                                                       Water for injections                                                                           0.40 l      (5.0 ml)                                         q.s. to                                                                       ______________________________________                                    

Doxorubicin.HCl (0.80g) was dissolved in 90 percent of the amount ofwater for injections, de-aerated by nitrogen bubbling. Ethanol was addedunder stirring and nitrogen bubbling. Hydrochloric acid 0.1 N was thenadded dropwise to adjust the pH of the solution to 3. De-aerated waterfor injections was finally added to bring the solution to its finalvolume (0.40 1).

The solution was filtered through a 0.22μ microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and at 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 12 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 8.

                                      TABLE 8                                     __________________________________________________________________________    INITIAL VALUES                                                                Concentration:1.979 mg/ml                                                                      pH = 3.11                                                    Relative % Assay: 100.0                                                       TEMPERATURE                                                                   4° C. 35° C.                                                                          45° C.                                                                         55° C.                                   TIME Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                                                              Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                       (weeks)                                                                            mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                                                              mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                       __________________________________________________________________________    1    2.010                                                                             101.6                                                                             1.965                                                                             99.3 1.947                                                                             98.4                                                                              1.750                                                                             88.4                                        2            1.957                                                                             98.9 1.910                                                                             96.5                                                                              1.645                                                                             83.1                                        3            1.895                                                                             95.8 1.737                                                                             87.8                                                                              1.356                                                                             68.5                                        4    1.927                                                                             97.3                                                                              1.818                                                                             91.9 1.678                                                                             84.8                                                12   1.939                                                                             97.9                                                                 __________________________________________________________________________     t.sub.90 (days) extrapolated according to Arrhenius equation:                 t.sub.90 at 4° C. = 1270 days                                          t.sub.90 at 8° C. = 780 days                                      

Similar stability data can be observed also for analogous solutionscontaining either doxorubicin hydrochloride at 5 mg/ml concentration, or4'-epi-doxorubicin, 4'-desoxy-doxorubicin,4'-desoxy-4'-iodo-doxorubicin, daunorubicin or 4-demethoxy-daunorubicin,as hydrochloride salts, at both 2 mg/ml and 5 mg/ml concentration.

EXAMPLE 9

    ______________________________________                                        Composition      for 80 vials                                                                              (for 1 vial)                                     ______________________________________                                        Doxorubicin.HCl  8.000 g     (100.00 mg)                                      N,N-Dimethylacetamide                                                                          0.060 l     (0.75 ml)                                        Propylene glycol 0.048 l     (0.60 ml)                                        Ethanol          0.012 l     (0.15 ml)                                        Hydrochloric acid 0.1N                                                                         pH = 3      (pH = 3)                                         q.s. to                                                                       Water for injections                                                                           0.400 l     (5.00 ml)                                        q.s. to                                                                       ______________________________________                                    

Doxorubicin.HCl (8.000 g) was dissolved in 90 percent of the amount ofwater for injections, de-aerated by nitrogen bubbling.N,N-dimethylacetamide, propylene glycol and ethanol were subsequentlyadded under stirring and nitrogen bubbling. The hydrochloric acid wasthen added dropwise to adjust the pH of the solution to 3. Furtherde-aerated water for injections was then added to bring the solution toits final volume (0.400 1).

The solution was filtered through a 0.22μ microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 8 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 9:

                                      TABLE 9                                     __________________________________________________________________________    INITIAL VALUES                                                                Concentration:20.07 mg/ml                                                                      pH = 2.99                                                    Relative % Assay: 100.0                                                       TEMPERATURE                                                                   4° C. 35° C.                                                                          45° C.                                                                         55° C.                                   TIME Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                                                              Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                       (weeks)                                                                            mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                                                              mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                       __________________________________________________________________________    1            19.14                                                                             95.4 17.81                                                                             88.7                                                                              14.84                                                                             73.9                                        2    19.97                                                                             99.5                                                                              19.07                                                                             95.0 16.27                                                                             81.1                                                                              12.36                                                                             61.6                                        3            18.08                                                                             90.1 14.62                                                                             72.9                                                                              10.04                                                                             50.0                                        4    20.06                                                                             99.9                                                                              18.03                                                                             89.8 13.20                                                                             65.8                                                8    19.69                                                                             98.1                                                                 __________________________________________________________________________     t.sub.90 (days) extrapolated according to Arrhenius equation:                 t.sub.90 at 4° C. = 846 days                                           t.sub.90 at 8° C. = 505 days                                      

Similar stability data can be observed for analogous solutionscontaining 4'-epi-doxorubicin or 4'-desoxy-doxorubicin, as hydrochloridesalts, at the same 20 mg/ml concentration.

EXAMPLE 10

    ______________________________________                                        Composition      for 80 vials                                                                              (for 1 vial)                                     ______________________________________                                        Doxorubicin.HCl  8.0 g       (100.0 mg)                                       Polyvinylpyrrolidone                                                                           20.0 g      (250.0 mg)                                       Hydrochloric acid 0.1N                                                                         pH = 3      (pH = 3)                                         q.s. to                                                                       Water for injections                                                                           0.4 l       (5.0 ml)                                         q.s. to                                                                       ______________________________________                                    

Doxorubicin.HCl (8.0 g) was dissolved in 90 percent of the amount ofwater for injections, de-aerated by nitrogen bubbling.Polyvinylpyrrolidone was added and dissolved under stirring and nitrogenbubbling. The hydrochloric acid was then added dropwise to adjust the pHof the solution to 3. Further de-aerated water for injections was thenadded to bring the solution to its final volume (0.4 1).

The solution was filtered through a 0.22μ microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45°C. and 35° C.) and 8 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 10:

                                      TABLE 10                                    __________________________________________________________________________    INITIAL VALUES                                                                Concentration: 19.57 mg/ml                                                                     pH = 2.62                                                    Relative % Assay: 100.0                                                       TEMPERATURE                                                                   4° C. 35° C.                                                                          45° C.                                                                         55° C.                                   TIME Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                                                              Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                       (weeks)                                                                            mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                                                              mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                       __________________________________________________________________________    1    19.54                                                                             99.9                                                                              19.11                                                                             97.6 16.88                                                                             86.2                                                                              12.48                                                                             63.8                                        2            18.43                                                                             94.2 14.13                                                                             72.2                                                                              6.00                                                                              30.7                                        3            18.02                                                                             92.1 11.57                                                                             59.1                                                                              2.61                                                                              13.3                                        4    19.58                                                                             100.1                                                                             17.36                                                                             88.7 9.23                                                                              47.2                                                8    19.34                                                                             98.8                                                                 __________________________________________________________________________     t.sub.90 (days) extrapolated according to Arrhenius equation:                 t.sub.90 at 4° C. = 2540 days                                          t.sub.90 at 8° C. = 1290 days                                     

Similar stability data can be observed for analogous solutionscontaining 4'-epi-doxorubicin or 4'-desoxy-doxorubicin, as hydrochloridesalts, at the same 20 mg/ml concentration.

EXAMPLE 11

    ______________________________________                                        Composition      for 80 vials                                                                              (for 1 vial)                                     ______________________________________                                        Doxorubicin.HCl  0.80 g      (10.0 mg)                                        N,N-Dimethylacetamide                                                                          0.12 l      (1.5 ml)                                         Hydrochloric acid 0.1N                                                                         pH = 3      (pH = 3)                                         q.s. to                                                                       Water for injections                                                                           0.40 l      (5.0 ml)                                         q.s. to                                                                       ______________________________________                                    

Doxorubicin.HCl (0.80 g) was dissolved in 90% of the amount of water forinjections, de-aerated by nitrogen bubbling. N,N-Dimethylacetamide wasadded under stirring and nitrogen bubbling. Hydrochloric acid 0.1N wasthen added dropwise to adjust the pH of the solution to 3. De-aeratedwater for injections was finally added to bring the solution to itsfinal volume (0.40 1).

The solution was filtered through a 0.22μ microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 8 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 11:

                                      TABLE 11                                    __________________________________________________________________________    INITIAL VALUES                                                                Concentration: 1.826 mg/ml                                                                     pH = 3.14                                                    Relative % Assay: 100.0                                                       TEMPERATURE                                                                   4° C. 35° C.                                                                          45° C.                                                                         55° C.                                   TIME Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                                                              Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                       (weeks)                                                                            mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                                                              mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                       __________________________________________________________________________    1    1.830                                                                             100.2                                                                             1.812                                                                             99.2 1.784                                                                             97.7                                                                              1.605                                                                             87.9                                        2    1.818                                                                             99.6                                                                              1.781                                                                             97.5 1.554                                                                             85.1                                                                              1.292                                                                             70.8                                        3            1.743                                                                             95.4 1.409                                                                             77.2                                                                              1.018                                                                             55.7                                        4    1.823                                                                             99.8                                                                              1.734                                                                             95.0 1.369                                                                             75.0                                                8    1.792                                                                             98.2                                                                 __________________________________________________________________________     t.sub.90 (days) extrapolated according to Arrhenius equation:                 t.sub.90 at 4° C. = 5815 days                                          t.sub.90 at 8° C. = 2920 days                                     

Similar stability data can be observed also for analogous solutionscontaining either doxorubicin hydrochloride at 5 mg/ml concentration, or4'-epi-doxorubicin, 4'-desoxy-doxorubicin,4'-epi-desoxy-4'-iodo-doxorubicin, daunorubicin or4-demethoxy-daunorubicin, as hydrochloride salts, at both 2 mg/ml and 5mg/ml concentration.

EXAMPLE 12

    ______________________________________                                        Composition      for 80 vials                                                                              (for 1 vial)                                     ______________________________________                                        Doxorubicin.HCl  0.80 g      (10.0 mg)                                        Propylene glycol 0.12 l      (1.5 ml)                                         Hydrochloric acid 0.1N                                                                         pH = 3      (pH = 3)                                         q.s. to                                                                       Water for injections                                                                           0.40 l      (5.0 ml)                                         q.s. to                                                                       ______________________________________                                    

Doxorubicin.HCl (0.80 g) was dissolved in 90% of the amount of water forinjections de-aerated by nitrogen bubbling. Propylene glycol was addedunder stirring and nitrogen bubbling. Hydrochloric acid 0.1 N was thenadded dropwise to adjust the pH of the solution to 3. De-aerated waterfor injections was finally added to bring the solution to its finalvolume (0.40 1).

The solution was filtered through a 0.22μ microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 4 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 12:

                                      TABLE 12                                    __________________________________________________________________________    INITIAL VALUES                                                                Concentration: 1.982 mg/ml                                                                     pH = 3.11                                                    Relative % Assay: 100.0                                                       TEMPERATURE                                                                   4° C. 35° C.                                                                          45° C.                                                                         55° C.                                   TIME Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                                                              Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                       (weeks)                                                                            mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                                                              mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                       __________________________________________________________________________    1    1.972                                                                             99.5                                                                              1.934                                                                             97.6 1.889                                                                             95.3                                                                              1.705                                                                             86.0                                        2            1.952                                                                             98.5 1.795                                                                             90.6                                                                              1.483                                                                             74.8                                        3            1.935                                                                             97.6 1.699                                                                             85.7                                                                              1.153                                                                             58.2                                        4    2.056                                                                             103.7                                                                             1.788                                                                             90.2 1.460                                                                             73.7                                                __________________________________________________________________________     t.sub.90 (days) extrapolated according to Arrhenius equation:                 t.sub.90 at 4° C. = 1794 days                                          t.sub.90 at 8° C. = 1025 days                                     

Similar stability data can be observed also for analogous solutionscontaining either doxorubicin hydrochloride at 5 mg/ml concentration, or4'-epi-doxorubicin, 4'-desoxy-doxorubicin,4'-desoxy-4'-iodo-doxorubicin, daunorubicin or 4-demethoxy-daunorubicin,as hydrochloride salts, at both 2 mg/ml 5 mg/ml concentration.

EXAMPLE 13

    ______________________________________                                        Composition      for 80 vials                                                                              (for 1 vial)                                     ______________________________________                                        Doxorubicin.HCl  0.80 g      (10.0 mg)                                        Polyethylene glycol 400                                                                        0.12 l      (1.5 ml)                                         Hydrochloric acid 0.1N                                                                         pH = 3      (pH = 3)                                         q.s. to                                                                       Water for injections                                                                           0.40 l      (5.0 ml)                                         q.s. to                                                                       ______________________________________                                    

Doxorubicin.HCl (0.80 g) was dissolved in 90% of the amount of water forinjections, de-aerated by nitrogen bubbling. Polyethylene glycol 400 wasadded under stirring and nitrogen bubbling. Hydrochloric acid 0.1 N wasthen added dropwise to adjust the pH of the solution to 3. De-aeratedwater for injections was finally added to bring the solution to itsfinal volume (0.40 1).

The solution was filtered through a 0.22μ microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 4 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 13:

                                      TABLE 13                                    __________________________________________________________________________    INITIAL VALUES                                                                Concentration: 1.907 mg/ml                                                                     pH = 3.07                                                    Relative % Assay: 100.0                                                       TEMPERATURE                                                                   4° C. 35° C.                                                                          45° C.                                                                         55° C.                                   TIME Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                                                              Conc.                                                                             Rel.%                                                                             Conc.                                                                             Rel.%                                       (weeks)                                                                            mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                                                              mg/ml                                                                             Assay                                                                             mg/ml                                                                             Assay                                       __________________________________________________________________________    1    1.871                                                                             98.1                                                                              1.797                                                                             94.2 1.668                                                                             87.5                                                                              1.484                                                                             77.8                                        2            1.710                                                                             89.7 1.608                                                                             84.3                                                                              1.237                                                                             64.9                                        3            1.739                                                                             91.2 1.551                                                                             81.3                                                                              1.007                                                                             52.8                                        4    1.873                                                                             98.2                                                                              1.693                                                                             88.8 1.453                                                                             76.2                                                __________________________________________________________________________     t.sub.90 (days) extrapolated according to Arrhenius equation:                 t.sub.90 at 4° C. = 1130 days                                          t.sub.90 at 8° C. = 680 days                                      

Similar stability data can be observed also for analogous solutionscontaining either doxorubicin hydrochloride at 5mg/ml concentration, or4'-epi-doxorubicin, 4'-desoxy-doxorubicin,4'-desoxy-4'-iodo-doxorubicin, daunorubicin or 4-demethoxy-daunorubicin,as hydrochloride salts, at both 2 mg/ml and 5 mg/ml concentration.

EXAMPLE 14

    ______________________________________                                        Composition      for 80 vials                                                                              (for 1 vial)                                     ______________________________________                                        Doxorubicin.HCl  0.8 g       (10 mg)                                          Hydrochloric acid 0.1N                                                                         pH = 3      (pH = 3)                                         q.s. to                                                                       Water for injections                                                                           0.4 l       (5 ml)                                           q.s. to                                                                       ______________________________________                                    

Doxorubicin.HCl (0.8 g) was dissolved in 90 percent of the amount ofwater for injections, de-aerated by nitrogen bubbling. The hydrochloricacid was then added dropwise to adjust the pH of the solution to 3.Further de-aerated water for injections was then added to bring thesolution to its final volume (0.4 1).

The solution was filtered through a 0.22μ microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 4° C. and 8° C. for up to 6 months.

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 14:

                  TABLE 14                                                        ______________________________________                                        INITIAL VALUES                                                                Concentration: 2.039 mg/ml                                                                          pH = 3.06                                               Relative % Assay: 100.0                                                               TEMPERATURE                                                                   4° C.      8° C.                                        TIME      Conc.   Rel.%       Conc. Rel.%                                     (months)  mg/ml   Assay       mg/ml Assay                                     ______________________________________                                        1         1.983   97.3        1.959 96.1                                      3         1.984   97.3        1.983 97.3                                      6         2.012   98.7        2.002 98.2                                      ______________________________________                                    

At the same conditions, similar stability data can be generally observedalso for the other solutions mentioned in the preceding examples.

We claim:
 1. A physiologically acceptable solution of anthracycline glycoside selected from the group consisting of idarubicin hydrochloride, doxorubicin hydrochloride and epirubicin hydrochloride dissolved in a physiologically acceptable aqueous solvent, having a pH adjusted to from 2.5 to 5.0 with a physiologically acceptable acid selected from the group consisting of hydrochloric acid, sulfuric acid, phosphoric acid, methane sulfonic acid, and tartaric acid, the concentration of said anthracycline glycoside being from 0.1 to 100 mg/ml, wherein said solution is contained in a sealed container.
 2. The solution of claim 1 wherein said physiologically acceptable aqueous solvent is selected from the group consisting of water, ethanol, polyethylene glycol, dimethylacetamide, and mixtures thereof.
 3. The solution of claim 1 wherein the physiologically acceptable aqueous solvent is water.
 4. The solution of claim 1 wherein the concentration of the doxorubicin is from 0.1 to 50 mg/ml.
 5. The solution of claim 4 wherein the concentration of doxorubicin is from 1 to 20 mg/ml.
 6. The solution of claim 1 wherein the concentration of doxorubicin is about 2 mg/ml.
 7. The solution of claim 1 wherein the concentration of doxorubicin is about 5 mg/ml.
 8. A sealed container containing a stable, intravenously injectable, sterile, pyrogen-free doxorubicin solution which consists essentially of doxorubicin hydrochloride dissolved in a physiologically acceptable solvent therefore, wherein said solution has a pH adjusted to 2.71 to 3.14 with a physiologically acceptable acid and has a concentration of doxorubicin of from 0.1 to 100 mg/ml.
 9. The anthracycline glycoside solution of claim 1, wherein said solution exhibits storage stability as a result of said pH being adjusted to the said pH range using said acids.
 10. The container of claim 8, wherein said doxorubicin solution exhibits storage stability as a result of said pH being adjusted to the said pH range using said acids.
 11. The solution of claim 1 wherein the concentration of anthracycline glycoside is about 1 mg/ml.
 12. The solution of claim 11 wherein the physiologically acceptable acid is hydrochloric acid.
 13. The solution of claim 12 wherein the anthracycline glycoside is idarubicin hydrochloride. 